CN111965231B - Semiconductor sensor for virus detection and preparation method and application thereof - Google Patents
Semiconductor sensor for virus detection and preparation method and application thereof Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/27—Association of two or more measuring systems or cells, each measuring a different parameter, where the measurement results may be either used independently, the systems or cells being physically associated, or combined to produce a value for a further parameter
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- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
Abstract
The invention belongs to the technical field of biochemical sensing, and discloses a semiconductor sensor for virus detection and a preparation method and application thereof, wherein the preparation method comprises the following steps: (1) preparing oxide or chalcogenide colloid semiconductor nano-materials; (2) coating a virus specific antigen or antibody on the surface of the colloidal semiconductor nano material to obtain a sensitive material; (3) based on the sensitive material, a virus detection sensor is prepared by adopting device structures such as a chemically modified electrode or a field effect transistor. According to the invention, the virus specific antigen or antibody is introduced to the surface of the colloidal semiconductor nano material, so that charge transfer caused by specific binding reaction between the virus antigen and the virus antibody is converted into a sensor electric signal through a semiconductor sensitive effect to be output, and the real-time performance and the convenience of a virus detection technology are improved.
Description
Technical Field
The invention belongs to the technical field of biochemical sensing, and particularly relates to a semiconductor sensor for virus detection and a preparation method and application thereof.
Background
The rapid, accurate and easily popularized virus detection technology has important significance for realizing early discovery, early isolation, early diagnosis and early treatment of virus infected people. Taking SARS-CoV-2 new coronavirus as an example, the virus causes infection in large-scale population due to its strong infectivity. Currently, the methods for detecting coronavirus are mainly classified into direct detection methods for viral nucleic acid and indirect detection methods for human serum antibody. Nucleic acid detection is one of the standard methods for determining viral infection, and is mainly implemented by obtaining a sample of upper respiratory tract secretions of a patient by a nasopharyngeal or oropharyngeal swab method and detecting viral ribonucleic acid (RNA) in the sample by utilizing a real-time fluorescent quantitative PCR (polymerase chain reaction) technology. The nucleic acid detection operation is complex, the detection time is long, and special requirements are provided for the skills of sampling and detection personnel and the biosafety qualification of a laboratory.
Compared with nucleic acid detection, the virus detection technology based on the specific binding reaction of the virus antigen and the antibody is quicker and more convenient, is suitable for various samples such as serum, saliva and the like, can reduce the requirements on detection personnel and laboratories, and obviously improves the detection efficiency. Taking the detection of antibodies against new coronavirus as an example, the most widely applied field detection means at present is a colloidal gold immunochromatographic kit, which mainly aims at detecting IgM (immunoglobulin M) and IgG (immunoglobulin G) antibodies in serum and urine samples and IgA (immunoglobulin A) antibodies in respiratory tract salivary mucosal secretions. The Chinese patent application discloses a new coronavirus antibody (IgG/IgM) colloidal gold immunochromatographic rapid detection test strip based on urine detection. And (3) directly dripping 2 drops of urine to be detected on the detection test paper, and judging the result by naked eyes within 15 minutes.
At present, the colloidal gold antibody detection technology mainly utilizes colloidal gold for color development and judges the result by naked eyes, has no quantitative detection capability and is easy to miss diagnosis or misjudge.
The technology for detecting various chemical and biological molecules by using the semiconductor sensor has the advantages of high sensitivity, low cost, convenience and quickness, and can improve the real-time property and convenience of detection. Meanwhile, the electrical signals can be displayed by numerical values, and the quantitative detection function is realized. Chinese patent CN103675034A discloses a semiconductor resistance type gas sensor using quantum dots as gas sensitive material and a preparation method thereof, wherein sodium nitrite short-chain ligand replaces long-chain organic ligand on the surface of quantum dots to realize low-concentration NO 2 High sensitivity detection of gas molecules at room temperature. The short-chain ligands on the surfaces of the colloid quantum dots have no specificity to gas molecules, do not participate in the reaction between the gas molecules and the quantum dots, and mainly have the effects of reducing surface long-chain organic ligands, exposing more gas molecule adsorption active sites, shortening the quantum dot spacing, improving carrier transport characteristics, and enabling the sensor performance to depend on the fact that the gas molecules directly perform adsorption reaction on atoms on the surfaces of the quantum dots to cause the change of the resistance value of the sensor.
The colloidal semiconductor nano material and the sensor thereof are mainly used for detecting gas micromolecules, and the colloidal semiconductor nano material for detecting biomacromolecules at present is mainly based on the fluorescence characteristic of colloidal quantum dots. The surface of the colloidal semiconductor nano material has rich active sites, the physicochemical property of the colloidal semiconductor nano material is easily influenced by the surface atomic composition and chemical reaction, and the colloidal semiconductor nano material has potential advantages in virus detection by combining the modification of virus specific antigen or antibody.
Disclosure of Invention
Aiming at the problems of long detection time, low convenience and difficulty in popularization of the existing virus detection method, the invention aims to provide a semiconductor sensor for virus detection and a preparation method and application thereof.
To achieve the above object, according to one aspect of the present invention, there is provided a method for manufacturing a semiconductor sensor for virus detection, comprising the steps of:
(1) preparing a colloidal semiconductor nano material;
(2) coating virus specific antigens or antibodies on the surface of the colloidal semiconductor nano material, wherein the virus antigens or antibodies can generate specific binding reaction on the virus marker of the target species, so as to obtain a sensitive material with the specific binding reaction on the virus marker of the target species;
(3) and coating the sensitive material on a chemical modification electrode, an interdigital electrode, a cantilever beam, a surface acoustic wave device or a field effect transistor electrode by adopting a drop coating, spin coating or spray printing film-forming process to construct a virus detection sensor.
As a further preferred aspect of the present invention, in the step (1), the colloidal semiconductor nanomaterial is specifically a colloidal quantum dot, nanowire, nanosheet, nanobelt of an oxide or chalcogenide, or a composite structure thereof, and is preferably a lead sulfide quantum dot, stannous sulfide quantum dot, zinc sulfide quantum dot, tin oxide quantum dot, tungsten oxide quantum dot, zinc oxide quantum dot, tin oxide nanowire, lead sulfide nanowire, bismuth sulfide nanobelt, molybdenum sulfide nanosheet, or tin sulfide nanosheet.
In a further preferred embodiment of the present invention, in the step (1), the colloidal semiconductor nanomaterial has a dimension of 1 to 100nm, and has a hydrophilic or hydrophobic ligand on the surface thereof, so that the colloidal semiconductor nanomaterial can be stably dispersed in water or an organic solvent.
As a further preferred aspect of the present invention, in the step (2), the surface of the colloidal semiconductor nanomaterial is coated with a virus antigen or antibody, specifically, by using a cross-linking method or a ligand exchange method, and the step is performed in a water-based solution formed by the colloidal semiconductor nanomaterial;
preferably, the crosslinking method is to modify mercaptopropionic acid, mercaptoethylamine or cysteine on the surface of the colloidal semiconductor nanomaterial to form a hydrophilic ligand with a carboxyl group or an amino group end capping, and then coat the virus antigen or antibody on the surface of the semiconductor nanomaterial by using glutaraldehyde, succinimide-4-cyclohexane-1-carbonate (SMCC) or 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) as a crosslinking agent; the ligand exchange method is that a virus antigen or antibody solution is directly mixed with a colloid semiconductor nano material in the solution for replacement reaction, so that the virus antigen or antibody directly replaces an organic ligand on the surface of the colloid semiconductor nano material to be coated on the surface of the semiconductor nano material.
As a further preferred aspect of the present invention, in step (2), surface blocking is performed after the viral antigen or antibody is coated, and further purification is performed;
preferably, the surface blocking step is to block the nonspecific protein binding sites on the surface of the semiconductor nano material and the surface of the cross-linking agent by using Bovine Serum Albumin (BSA);
preferably, in the purification treatment step, the sample is concentrated and purified 3-5 times by using an ultrafiltration centrifugal tube, and after centrifugal washing, the final product is redissolved in a phosphate buffer solution.
According to still another aspect of the present invention, there is provided a method for manufacturing a semiconductor sensor for virus detection, comprising the steps of:
(1) preparing a colloidal semiconductor nano material;
(2) coating the colloid semiconductor material on a chemical modification electrode, an interdigital electrode, a cantilever beam, a surface acoustic wave device or a field effect transistor electrode by adopting a dripping, spin coating or spraying printing film-forming process to form a semiconductor nano film;
(3) and virus specific antigens or antibodies are coated on the surface of the semiconductor nano film, and the virus antigens or antibodies can generate specific binding reaction on the virus marker of the target species, so that the semiconductor sensor with specific recognition on the virus marker of the target species is obtained.
As a further preferred aspect of the present invention, in the step (3), the surface of the semiconductor nanomaterial is coated with a viral antigen or antibody specifically by a cross-linking method or a ligand exchange method;
preferably, the crosslinking method is to modify mercaptopropionic acid, mercaptoethylamine or cysteine on the surface of the semiconductor nano-film to form a hydrophilic ligand with a carboxyl group or an amino group end capping, and then to coat the virus antigen or the antibody on the surface of the semiconductor nano-film by using glutaraldehyde, succinimide-4-cyclohexane-1-carbonate (SMCC) or 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) as a crosslinking agent; the ligand exchange method is to drop virus antigen or antibody solution on the surface of the film for replacement reaction, so that the virus antigen or antibody directly replaces organic ligands on the surface of the colloidal semiconductor nano film to coat the surface of the semiconductor nano film.
Preferably, in the step (3), surface blocking is performed after the viral antigen or antibody is coated; the surface blocking step is to block the nonspecific protein binding sites on the surface of the semiconductor nano-film and the surface of the cross-linking agent by adopting Bovine Serum Albumin (BSA).
According to another aspect of the present invention, there is provided a virus detection sensor prepared by the above preparation method.
According to another aspect of the invention, the invention provides the application of the virus detection sensor in a hand-held virus detector or an intelligent terminal; preferably, the intelligent terminal is an intelligent mobile phone;
more preferably, the virus detector or the intelligent terminal is used for detecting SARS-CoV-2 novel coronavirus.
Compared with the prior art, the technical scheme of the invention has the advantages that the existing colloidal gold test strip technology depends on a double-antibody sandwich method, the pairing design of two antibodies or two antigens is a difficult point, the antigens and the screened antibodies need to be designed aiming at different epitopes of the target protein, and the semiconductor sensor detection scheme only needs one antigen or antibody, so that the technical process can be simplified, and the production cost can be reduced. The invention realizes the detection of virus by utilizing the combination of the surface antigen or antibody of the colloidal semiconductor nano material and the virus marker, the colloidal semiconductor nano material has large specific surface area and rich surface activity stable points, the physicochemical property is easily influenced by the surface atomic composition and chemical reaction, the specific combination reaction of the virus antibody and the antigen can cause the obvious change of the electrical signal of the semiconductor nano material, which is beneficial to improving the specificity and the sensitivity of the detection result, simultaneously, the structure of the interdigital electrode of the chemical modification electrode, the cantilever beam, the surface acoustic wave device and the field effect transistor is adopted to realize the signal conversion and the amplification, because the semiconductor sensor adopts the electric signal for reading, compared with the prior immunochromatography technology, the detection speed is higher by depending on the naked eye judgment, the detection sensitivity and the reliability are higher, the in-situ cooperative detection of the new coronavirus antigen and the antibody can be realized at the same time and the same place, has the characteristics of higher accuracy, rapidness, high efficiency and easy popularization. In addition, the sensor is small in size, electrical signals are easy to integrate with a signal transmission device, and meanwhile, the sensor based on the cantilever beam and the surface acoustic wave device has the advantages of being wireless and passive, can read detection signals remotely, and improves the reliability and convenience of virus detection. The virus detection method can realize non-contact test between medical staff and an infected person in the virus detection process, greatly ensures the detection safety, and can be popularized and applied to scenes of quick screening, large-scale investigation, clinical diagnosis, epidemic investigation, vaccine development and the like of new coronavirus carriers in communities, hospitals and public places.
When the virus sensor device is applied, a serum, saliva or urine sample can be added into a phosphate buffer solution, a mixed solution is immediately dripped onto the sensor, the signal response of the sensor is tested within 1-5 minutes, and a detection result is obtained through the signal response. The virus marker of the target species in the invention can be SARS-CoV-2 novel coronavirus specific antibody or antigen, or influenza virus antibody or antigen; the target virus for which the present invention is applicable may be, in particular, SARS-CoV-2 novel coronavirus. As the biological materials such as antigens and antibodies of the present invention, commercially available products such as novel coronavirus antigen (ATMP02479COV (RBD)) and novel coronavirus antibody (ATMA10176Mo) produced by Projia organism (Wuhan) science and technology Limited can be used.
Drawings
FIG. 1 is a schematic diagram of colloidal quantum dots surface-modified specific virus antigen detection virus antibodies.
FIG. 2 is the results of the detection of new coronavirus antibodies using integrated pulsed voltammetry (DPV) mode with planar three electrodes in example 1. The curve marked by delta PBS is a DPV curve of the lead sulfide modified electrochemical sensor in Phosphate (PBS) buffer solution, the curve marked by O antibody is a DPV curve of the electrochemical sensor added with 1 muL of new coronavirus standard antibody in the phosphate buffer solution, and the graph shows that after the antibody is added, the current is obviously increased, which indicates that the specific binding reaction of the new coronavirus antigen and the antibody causes the increase of the current peak value.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
(1) Preparing the lead sulfide (PbS) quantum dot material. Specifically, 1.8g of lead oxide, 6mL of OA, and 20mL of Octadecene (ODE) were stirred at a high speed in a three-neck flask, and the temperature of the precursor was raised to 120 ℃ after evacuation. The sulfur precursor was prepared by dissolving 280. mu.L of bis (trimethylsilyl) sulfide in 10ml of ODE evacuated in a glove box using a pipette gun. And rapidly injecting a sulfur precursor into the lead precursor in a nitrogen environment, reacting for 30s, and then placing into a cold water bath for rapid cooling. And collecting the precipitate, washing the precipitate for a plurality of times by using toluene acetone, centrifugally collecting the precipitate, and drying the precipitate in vacuum to obtain the PbS quantum dot.
(2) And dispersing the PbS quantum dots in n-octane to obtain a PbS colloidal quantum dot solution with the concentration of 10-50mg/mL, dripping 0.5 mu L of the PbS quantum dots onto a working electrode of a planar three-electrode by using a pipette, coating for 100s at the rotating speed of 260rpm in a spin coating film forming mode, repeating the step for 4 times, and drying for 10 minutes at room temperature to obtain the lead sulfide quantum dot modified planar three-electrode.
(3) And dripping and coating 1 mu L of commercial standard antigen on the surface of the lead sulfide quantum dot film, and carrying out antigen incubation treatment for 1 hour at 37 ℃. And after treatment, washing the surface of the film by adopting a phosphate buffer solution, and removing residual substances on the surface to obtain the antigen-coated lead sulfide quantum dot film. And (2) dropwise adding 200 mu L of bovine serum albumin solution (10 mg/mL) and wrapping the solution on the surface of the antigen-coated lead sulfide quantum dot film, soaking for 2 hours at room temperature to seal the surface of the quantum dot, then washing the surface of the film by using phosphate buffer solution, and removing residual substances on the surface to obtain the bovine serum albumin-sealed lead sulfide quantum dot film.
(4) 200 mu L of phosphate buffer solution is dripped on the surface of the planar three-electrode, and an electrochemical workstation is adopted to test the cyclic voltammetry characteristic, the integral pulse voltammetry characteristic and the electrochemical impedance spectrum of the surface of the working electrode within 1-5 minutes. And then injecting antibody solutions with different concentrations into the 200 mu L of phosphate buffer solution, respectively testing cyclic voltammetry characteristics, integrated pulse voltammetry characteristics and electrochemical impedance spectroscopy characteristics, and judging the antibody concentration by recording the magnitude of the oxidation reduction peak current and the magnitude of impedance of the electrochemical test.
Example 2
(1) Preparing the stannous sulfide (SnS) quantum dot material. Specifically, 1mM stannous chloride (SnCl) is taken 2 ) 5mL of oleic acid and 5mL of octadecene are stirred at high speed in a three-neck flask, heated to 120 ℃, vacuumized for 30 minutes, and subjected to nitrogen atmosphere for 6 minutes -1 The temperature is raised to 150 ℃ at the temperature raising rate, 0.5mL of 1-dodecanethiol (DDT) is injected, the heating is continued to 180 ℃, and 1mM thioacetamide is dissolved in 2mL of oleylamine to prepare a precursor of sulfur. Under nitrogenAnd rapidly injecting the sulfur precursor into the tin precursor at 180 ℃ under the environment, reacting for 30min, and cooling to room temperature. Washing with ethanol for several times, centrifuging, collecting precipitate, and vacuum drying to obtain SnS quantum dots.
(2) Selecting a phosphate buffer solution, adding SnS quantum dots, adding 3-mercaptopropionic acid (3-MPA) to replace organic ligands on the surfaces of the quantum dots, then coating antigens capable of specifically detecting virus antibodies on the surfaces of the quantum dots by using EDC as a cross-linking agent, and dissolving the final product in the phosphate buffer solution after centrifugal washing.
(3) An electrofluid spray printing film forming process is selected, SnS quantum dot solution modified with specific antigen is coated on a grid electrode of a High Electron Mobility Transistor (HEMT), drying is carried out at room temperature to obtain a quantum dot sensitized transistor device, and a semiconductor parameter tester is adopted to record the output characteristics of the transistor.
(4) Adding 20 mu L of serum sample into 100 mu L of phosphate buffer solution, immediately dripping 50 mu L of phosphate buffer solution on the quantum dot sensitized field effect transistor sensor, testing the output characteristic of the transistor within 1-5 minutes, and comparing the leakage current (I) of the transistor D ) The concentration of the virus antibody is judged according to the change size.
In the above embodiment, only three planar electrodes and a field effect transistor are taken as examples, and the present invention may also adopt sensor structures such as a surface acoustic wave device, a resonant cantilever beam, and an interdigital electrode to implement signal conversion and amplification. In addition to the above-mentioned synthesis process, the colloidal semiconductor nanomaterial can be synthesized by other synthesis methods known in the art, and of course, commercially available colloidal semiconductor nanomaterials can also be used directly. The biological materials such as antigens and antibodies mentioned in this patent can be commercially available, such as new coronavirus antigen (ATMP02479COV (RBD)) and new coronavirus antibody (ATMA10176Mo) produced by Probiotics (Wuhan science and technology Limited.
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A method for manufacturing a semiconductor sensor for virus detection, comprising the steps of:
(1) preparing a colloidal semiconductor nano material; the dimension of the colloidal semiconductor nano material is 1-100 nm, and a hydrophilic or hydrophobic ligand is arranged on the surface of the colloidal semiconductor nano material and can be stably dispersed in water or an organic solvent;
(2) coating virus specific antigens or antibodies on the surface of the colloidal semiconductor nano material, wherein the virus antigens or antibodies can generate specific binding reaction on the virus marker of the target species, so as to obtain a sensitive material with the specific binding reaction on the virus marker of the target species;
(3) coating the sensitive material on a chemical modification electrode, an interdigital electrode, a cantilever beam, a surface acoustic wave device or a field effect transistor grid by adopting a drop coating, spin coating or spray printing film-forming process to construct a virus detection sensor;
in the step (2), the surface of the colloidal semiconductor nano material is coated with virus antigen or antibody, specifically by adopting a cross-linking method or a ligand exchange method, and the step is completed in a water-based solution formed by the colloidal semiconductor nano material;
the crosslinking method specifically comprises the steps of modifying mercaptopropionic acid, mercaptoethylamine or cysteine on the surface of a colloidal semiconductor nano material to form a hydrophilic ligand with a carboxyl or amino end capping, and then coating a virus antigen or an antibody on the surface of the semiconductor nano material by using glutaraldehyde, succinimide-4-cyclohexane-1-carbonate or 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride as a crosslinking agent; the ligand exchange method is that a virus antigen or antibody solution is directly mixed with a colloid semiconductor nano material in the solution for replacement reaction, so that the virus antigen or antibody directly replaces an organic ligand on the surface of the colloid semiconductor nano material to be coated on the surface of the semiconductor nano material.
2. The preparation method according to claim 1, wherein in the step (1), the matrix semiconductor nanomaterial in the colloidal semiconductor nanomaterial is a colloidal quantum dot, a nanowire, a nanosheet, a nanobelt of an oxide or a chalcogenide, or a composite structure thereof.
3. The preparation method according to claim 2, wherein in the step (1), the base semiconductor nanomaterial in the colloidal semiconductor nanomaterial is specifically a lead sulfide quantum dot, a stannous sulfide quantum dot, a zinc sulfide quantum dot, a tin oxide quantum dot, a tungsten oxide quantum dot, a zinc oxide quantum dot, a tin oxide nanowire, a lead sulfide nanowire, a bismuth sulfide nanobelt, a molybdenum sulfide nanosheet, or a tin sulfide nanosheet.
4. The method according to claim 1, wherein in the step (2), surface blocking is performed after the viral antigen or antibody is coated, and further purification is performed;
wherein, the surface blocking step is to block the nonspecific protein binding sites on the surface of the semiconductor nano material and the surface of the cross-linking agent by adopting bovine serum albumin;
in the purification treatment step, the sample is concentrated and purified for 3-5 times by using an ultrafiltration centrifugal tube, and the final product is redissolved in a phosphate buffer solution after centrifugal washing.
5. A method for manufacturing a semiconductor sensor for virus detection, comprising the steps of:
(1) preparing a colloidal semiconductor nano material; the dimension of the colloidal semiconductor nano material is 1-100 nm, and a hydrophilic or hydrophobic ligand is arranged on the surface of the colloidal semiconductor nano material and can be stably dispersed in water or an organic solvent;
(2) coating the colloid semiconductor material on a chemical modification electrode, an interdigital electrode, a cantilever beam, a surface acoustic wave device or a field effect transistor grid to form a semiconductor nano film by adopting a dripping, spin coating or spraying printing film-forming process;
(3) coating virus specific antigens or antibodies on the surface of the semiconductor nano film, wherein the virus antigens or antibodies can generate specific binding reaction on the virus markers of the target species, so as to obtain the semiconductor sensor with specific recognition on the virus markers of the target species;
in the step (3), the surface of the semiconductor nano material is coated with the virus antigen or antibody by adopting a cross-linking method or a ligand exchange method;
the crosslinking method specifically comprises the steps of modifying mercaptopropionic acid, mercaptoethylamine or cysteine on the surface of the semiconductor nano-film to form a hydrophilic ligand with a carboxyl or amino end capping, and then coating a virus antigen or an antibody on the surface of the semiconductor nano-film by using glutaraldehyde, succinimide-4-cyclohexane-1-carbonate or 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride as a crosslinking agent; the ligand exchange method is to drop virus antigen or antibody solution on the surface of the film for replacement reaction, so that the virus antigen or antibody directly replaces organic ligands on the surface of the colloidal semiconductor nano material to coat the surface of the semiconductor nano film.
6. The method according to claim 5, wherein in the step (3), surface blocking is further performed after the viral antigen or antibody is coated;
wherein, the surface blocking step is to block the nonspecific protein binding sites on the surface of the semiconductor nano-film and the surface of the cross-linking agent by adopting bovine serum albumin.
7. A virus detection sensor produced by the production method according to any one of claims 1 to 6.
8. Use of the virus detection sensor according to claim 7 in the manufacture of a hand-held virus detector or integrated into an intelligent terminal.
9. The application of claim 8, wherein the smart terminal is a smart phone.
10. The use of claim 8, wherein the hand-held virus detector or intelligent terminal is capable of being used for the novel coronavirus detection of SARS-CoV-2.
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| CN114813877B (en) * | 2022-05-31 | 2023-03-14 | 华中科技大学 | Sensor for detecting glucose, preparation method and application thereof |
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